H2S调节miR-21表达抑制内质网应激介导肺成纤维化细胞凋亡

目的：探讨H2S是否可调节miR-21表达抑制内质网应激介导肺成纤维细胞凋亡。方法：体外培养小鼠成纤维细胞(NIH3T3)，随机分为对照组、TGF-β1组和NaHS干预组。采用CCK-8分别检测细胞增殖，流式细胞仪检测NIH3T3凋亡率；qRT-PCR法和Western blot分别分析葡萄糖调节蛋白78(GRP78)、增强子结合蛋白同源蛋白(CHOP)和caspase-12的表达；检测miR-21是否参与H2S抑制ERS介导的肺成纤维化细胞凋亡,将其随机分为3组：对照组、miR-21激动剂(miR-21 agomir)组和miR-21拮抗剂(miR-21 antagomir)组。miR-21激动剂组和miR-21拮抗剂组分别给予40 μmol/L的NaHS 预处理，再进行TGF-β1处理，检测GRP78、CHOP 及caspase-12的表达。结果：与对照组比较，NaHS干预组小鼠肺成纤维化细胞的增殖率、细胞凋亡率、miR-21蛋白及mRNA的表达均明显降低；与TGF-β1组比较，NaHS可明显降低内质网应激相关因子GRP78、CHOP及 caspase-12蛋白和mRNA表达；与阴性对照组比较，miR-21 agomir组GRP78、CHOP 及caspase-12的蛋白及mRNA表达均显著升高。而与miR-21 agomir组比较，miR-21 antagomir组结果则与之相反。结论：H2S可通过调节miR-21的表达，调控TGF-β1诱导小鼠肺成纤维化细胞的ERS稳态减少细胞凋亡，发挥其内源性的保护作用。

Hydrogen sulfide inhibits endoplasmic reticulum stress mediated apoptosis of pulmonary fibroblasts by regulating expression of microRNA-21

Objective:To explore whether H2S regulating the expression of miR 21 and then inhibition endoplasmic reticulum stress-mediated apoptosis of pulmonary fibroblasts.Methods: Mice pulmonary fibroblasts NIH3T3 were cultured in vitro.Then the cells were divided into a control group,a TGF-β1 group,and an H2S protection group.The rate of cell proliferation was monitored by CCK-8,and the apoptosis rate of the pulmonary fibroblasts by flow cytometry.The mRNA and protein expressions of GRP78,CHOP and caspase-12 were monitored by real-time RT-PCR and Western blot.To verify whether miR-21 was involved in the ERS-mediated apoptosis of the pulmonary fibroblasts,these cells were divided into three groups in random including the control group,miR-21 agomir group and miR-21 antagomir group.Both miR-21 agomir and miR-21 antagomir group were subjected to pretreat with NaHS at 40 μmol/L at 60 min,whereafter receive TGF-β1 after transfected,followed by detection of the expression of GRP78,CHOP and caspase-12. Results: Compared with the control,the rate of cell proliferation and apoptosis significantly reduced in the H2S protection group,similarly to the expression of miR-21.Compared with the TGF-β1 group,NaHS could obviously reduce the expression of GRP78,caspase-12 and CHOP.Compared with the negative group,the expression of GRP78，CHOP and caspase-12 significantly upregulated in miR-21 agomir groupHowever,the results was opposite in the miR-21 antagomir group. Conclusion: H2S may inhibit pulmonary fibroblasts by regulating the expression of miR-21 and reducing ERS-mediated cell apoptosis.